S-Adenosylmethionine (SAM) is a ubiquitous biomolecule serving mainly as a methyl donor. Our recent studies revealed that SAM controls antibiotic production in Streptomyces. In this study, the functional mode of SAM was studied in S. coelicolor and S. antibioticus ATCC11891, employing S-adenosylhomocysteine (SAH), a methylation reaction product of SAM. Actinorhodin biosynthesis did not require SAM as a methyl donor, whereas SAH enhanced the actinorhodin biosynthesis up to the level comparable to SAM, and the most effective concentration of SAH was higher than that of SAM. In the case of oleandomycin that requires SAM for its biosynthesis, both SAM and SAH at the concentration as low as 100 mM showed comparable efficacy in enhancing the production; SAM at 1 mM concentration additionally stimulated to give a 5-fold enhancement of oleandomycin production. In vitro autophosphorylation of protein kinase AfsK was found to be activated by both SAM and SAH, as well as other structurally related compounds. Our studies demonstrate that SAM regulates antibiotic biosynthesis in a mode independent of its role as a methyl donor and suggest that SAM acts directly as an intracellular signaling molecule for Streptomyces.
Pranaw, Kumar;Singh, Surender;Dutta, Debjani;Singh, Nirpendra;Sharma, Garima;Ganguly, Sudershan;Kalia, Vinay;Nain, Lata
Journal of Microbiology and Biotechnology
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v.23
no.11
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pp.1536-1543
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2013
Proteases produced by Xenorhabdus are known to play a significant role in virulence leading to insect mortality. The present study was undertaken to purify and characterize protease from Xenorhabdus indica, an endosymbiont of nematode Steinernema thermophilum, and to decipher its role in insect mortality and its efficacy to control Helicoverpa armigera. A set of 10 strains of Xenorhabdus isolated from different regions of India were screened for protease activity on the basis of zone of clearing on gelatin agar plates. One potent strain of Xenorhabdus indica was selected for the production of protease, and the highest production (1,552 U/ml) was observed at 15-18 h of incubation at $28^{\circ}C$ in soya casein digest broth. The extracellular protease was purified from culture supernatant using ammonium sulfate precipitation and ion-exchange chromatography. The enzyme was further characterized by SDS-PAGE and zymography, which confirmed the purity of the protein and its molecular mass was found to be ~52 kDa. Further MALDI-TOF/TOF analysis and effect of metal chelating agent 1,10-phenanthrolin study revealed the nature of the purified protease as a secreted alkaline metalloprotease. The bioefficacy of the purified protease was also tested against cotton bollworm (Helicoverpa armigera) and resulted in $67.9{\pm}0.64%$ mortality within one week. This purified protease has the potential to be developed as a natural insecticidal agent against a broad range of agriculturally important insects.
Choi, Noorie;Chang, Ji Hyun;Kim, Suzy;Kim, Hak Jae
Radiation Oncology Journal
/
v.35
no.2
/
pp.144-152
/
2017
Purpose: The role of radiotherapy (RT) was largely deserted after the introduction of platinum-based chemotherapy, but still survival rates are disappointingly low. This study focuses on assessing the clinical efficacy of RT in relation to chemotherapy resistance. Materials and Methods: From October 2002 to January 2015, 44 patients were diagnosed with epithelial ovarian cancer (EOC) and treated with palliative RT for persistent or recurrent EOC. All patients received initial treatment with optimal debulking surgery and adjuvant platinum-based chemotherapy. The biologically effective dose (BED) was calculated with ${\alpha}/{\beta}$ set at 10. Ninety-four sites were treated with RT with a median BED of 50.7 Gy (range 28.0 to 79.2 Gy). The primary end-point was the in-field local control (LC) interval, defined as the time interval from the date RT was completed to the date any progressive or newly recurring disease within the RT field was detected on radiographic imaging. Results: The median follow-up duration was 52.3 months (range 7.7 to 179.0 months). The 1-year and 2-year in-field LC rates were 66.0% and 55.0%, respectively. Comparisons of percent change of in-field tumor response showed similar distribution of responses among chemoresistant and chemosensitive tumors. On multivariate analysis of predictive factors for in-field LC analyzed by sites treated, $BED{\geq}50Gy$ (hazard ratio, 0.4; confidence interval, 0.2-0.9; p = 0.025) showed better outcomes. Conclusion: Regardless of resistance to platinum-based chemotherapy, RT can be a feasible treatment modality for patients with persistent of recurrent EOC. The specific role of RT using updated approaches needs to be reassessed.
One hundred & seventy four consecutive free-flap transfers were reviewed to analyze distribution of the type of reconstructions, kinds of donor flaps as well incidence of complications. The role of emergent exploration and the effect of preoperative wound conditions in flap survival were evaluated. Free flap transfer for head and neck reconstruction was most common as 93 cases, followed by for upper extremity of 30 cases, for lower extremity 30 cases, 18 penile reconstructions and for trunk & breast 3 cases. Nine flaps exhibited signs of ciruclatory insufficiency between 5 hours and 7 days. Three were managed conservatively with ultimate partial necrosis of the flaps. Eight flaps required return to the operating room. On exploration, early arterial occlusion was revealed in 1 flap, late arterial occlusion in 2 flaps, early venous occlusion in 1 flap, late venous thrombosis in 2 flaps, prolonged venous spasm in 1 and hematoma in 1 flap. The average time from the first abnormal examination to exploration was 2.6 hours. There were no false-positive explorations. Four free flaps failed in spite of the correction of the cause of circulatory compromise. The remaining 4 flaps were salvaged following the correction the casuse. Recipient vessel problems such as irradiation and infection were the most common cause of circulatory crisis. Among the eight flaps requiring return to the operating room, single vein was anastomosed in three flaps and two veins in the remaining five. In the totally failed four flaps only single vein was anastomosed in three cases. The results of this study demonstrate the efficacy of clinical monitoring and the role of early exploration. Precautious selection of recipient vessels and two vein anastomosis are recommended for safe and better prognosis.
Background: Oral squamous cell carcinoma (OSCC) is an important malignancy throughout the world; early detection is an important criterion for achieving high cure rate. Out of the many reported markers for OSCC, this study validated the efficacy of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in differentially diagnosing premalignant oral lesions and OSCC. Also, the study aimed to correlate the levels of salivary and serum TNF-${\alpha}$ with clinicopathologic factors. Materials and Methods: A prospective experimental laboratory study was designed. Serum and salivary samples from 100 subjects in each group of healthy control, premalignant disease (PMD) and OSCC were collected for the study following appropriate exclusion and inclusion criteria. Serum and salivary level of TNF-${\alpha}$ was analysed by enzyme linked immunosorbent assay. The data obtained were subjected to appropriate statistical analysis. Results: Increased level of both serum and salivary TNF-${\alpha}$ was observed in OSCC subjects compared to healthy control and PMD group. Receiver operator characteristic curve analysis and area under curve values showed high specificity and sensitivity for salivary TNF-${\alpha}$ in differentiating OSCC from PMD and healthy controls. There was significant increase in TNF-${\alpha}$ level in moderately and poorly differentiated lesion compared to well differentiated lesion and in stage IV of clinical stage. A positive correlation was observed only with histological grading of OSCC and TNF-${\alpha}$. Conclusions: Salivary TNF-${\alpha}$ is proved to be superior for detecting OSCC. Increase in TNF-${\alpha}$ with histological grading and clinical staging suggests a role in prognosis.
The structures and processes of medical education have changed little since the publication of Flexner's report, which stressed the scientific orientation of medical education and the curricular structure of 2 years of formal knowledge education and 2 years of clinical experience. However, the previous perspectives on medical education are facing challenges, and these call for new pedagogy and theories on which to base medical education practice. Considering that social dimensions of learning have been emphasized in practice, perspectives that integrate these aspects are needed. Among the various learning theories, social cognitive theory refers to the theoretical framework which contends that learning occurs within interactions with others and environments. From a social cognitive standpoint, learning through observation is a critical component in human functioning. Indeed, observational learning has particular significance in medical education in that it provides the context for which the importance and meaning of role models can be understood. In addition, as theoretical constructs such as self-efficacy and outcome expectations allow us to establish an effective learning environment, exploring the concepts of the theory could be beneficial to medical education practice. In this context, the present review article aims to provide a glimpse of the fundamental assumptions and theoretical concepts of social cognitive theory and discusses the implications the theory has on teaching and learning. Further, a review of previous studies could help explain how the theory has informed medical education practice. Finally, the author will conclude with the implications and limitations of applying social cognitive theory in medical education.
Musa, Dickson A.;Aremu, Kolawole H.;Ajayi, Abraham;Smith, Stella I.
Microbiology and Biotechnology Letters
/
v.48
no.2
/
pp.230-235
/
2020
The global evolution of antibiotic resistance has threatened the efficacy of available treatment options with ravaging impacts observed in developing countries. As a result, investigations into the prevalence of antibiotic resistance and the role of plasmids are crucial. In this study, we investigated the presence and distribution of blaTEM and gyrA genes, plasmid profiles, and the antimicrobial susceptibility pattern of Salmonella strains isolated from raw meat and stool sources across Niger State, Nigeria. Ninety-eight samples, comprising 72 raw meat and 26 stool samples, were screened for Salmonella spp. The antimicrobial susceptibility of Salmonella isolates to 10 commonly used antimicrobial agents was determined using the KirbyBauer disc diffusion method. Isolates were further analyzed for plasmids, in addition to PCR amplification of beta-lactamase (blaTEM) and gyrA genes. A total of 31 Salmonella spp. were isolated, with 22 from raw meat (70.97%) and 9 from stool (29.03%). Salmonella spp. with multiple resistance patterns to ceftazidime, cefuroxime, ceftriaxone, erythromycin, ampicillin, cloxacillin, and gentamicin were detected. Ofloxacin and ciprofloxacin were found to be the most effective among the antibiotics tested, with 67.7% and 93.5% susceptible isolates, respectively. Nine (29.03%) isolates harbored plasmids with molecular sizes ranging between 6557 bp and 23137 bp. PCR amplification of gyrA was detected in 1 (3.23%) of the 31 isolates while 28 isolates (90.32%) were positive for blaTEM. This study shows the incidence of antibiotic resistance in Salmonella isolates and the possible role of plasmids; it also highlights the prevalence of ampicillin resistance in this local population.
Kim, You-Sun;Hong, Goohyeon;Kim, Doh Hyung;Kim, Young Min;Kim, Yoon-Keun;Oh, Yeon-Mok;Jee, Young-Koo
Experimental and Molecular Medicine
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v.50
no.11
/
pp.9.1-9.10
/
2018
Although the positive effects of recombinant fibroblast growth factor-2 (rFGF-2) in chronic obstructive pulmonary disease (COPD) have been implicated in previous studies, knowledge of its role in COPD remains limited. The mechanism of FGF2 in a COPD mouse model and the therapeutic potential of rFGF-2 were investigated in COPD. The mechanism and protective effects of rFGF-2 were evaluated in cigarette smoke-exposed or elastase-induced COPD animal models. Inflammation was assessed in alveolar cells and lung tissues from mice. FGF-2 was decreased in the lungs of cigarette smoke-exposed mice. Intranasal use of rFGF-2 significantly reduced macrophage-dominant inflammation and alveolar destruction in the lungs. In the elastase-induced emphysema model, rFGF-2 improved regeneration of the lungs. In humans, plasma FGF-2 was decreased significantly in COPD compared with normal subjects (10 subjects, P = 0.037). The safety and efficacy of inhaled rFGF-2 use was examined in COPD patients, along with changes in respiratory symptoms and pulmonary function. A 2-week treatment with inhaled rFGF-2 in COPD (n = 6) resulted in significantly improved respiratory symptoms compared with baseline levels (P < 0.05); however, the results were not significant compared with the placebo. The pulmonary function test results of COPD improved numerically compared with those in the placebo, but the difference was not statistically significant. No serious adverse events occurred during treatment with inhaled rFGF-2. The loss of FGF-2 production is an important mechanism in the development of COPD. Inhaling rFGF-2 may be a new therapeutic option for patients with COPD because rFGF-2 decreases inflammation in lungs exposed to cigarette smoke.
Background: The efficacy of ginseng, the representative product of Korea, and its chemical effects have been well investigated. The ginsenoside RG3 has been reported to exhibit apoptotic, anticancer, and antidepressant-like effects. Methods: In this report, the putative effect of RG3 on several cellular function including cell survival, differentiation, development and aging process were evaluated by monitoring each specific marker. Also, mitochondrial morphology and function were investigated in ultraviolet (UV)-irradiated normal human dermal fibroblast cells. Results: RG3 treatment increased the expression of extracellular matrix proteins, growth-associated immediate-early genes, and cell proliferation genes in UV-irradiated normal human dermal fibroblast cells. And, RG3 also resulted in enhanced expression of antioxidant proteins such as nuclear factor erythroid 2-related factor-2 and heme oxygenase-1. In addition, RG3 affects the morphology of UV-induced mitochondria and plays a role in protecting mitochondrial dysfunction. Conclusioin: RG3 restores mitochondrial adenosine triphosphate (ATP) and membrane potential via its antioxidant effects in skin cells damaged by UV irradiation, leading to an increase in proteins linked with the extracellular matrix, cell proliferation, and antioxidant activity.
Kim, Wangin;Kim, Jieun;Lee, Seonhee;Moon, Yangsun;Lee, Sukhee;Park, Sooyun;Na, Changsu
The Korea Journal of Herbology
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v.28
no.3
/
pp.45-51
/
2013
Objectives : This study was conducted to measure the antioxidative and antimicrobial activities of water- and ethanol-extracts from Betula platyphylla var. japonica, Punica granatum and Rhus javanica against various species of anaerobic bacteria. Methods : In order to evaluate the antioxidative and antimicrobial activities of water- and ethanol-extracts, DPPH radical scavenging and superoxide dismutase (SOD)-like activities were measured in various species such as Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Propionibacterium acnes that induce skin inflammation. Also the total amount of phenol in each water- and ethanol-extract was measured to identify its role in the antioxidative and antimicrobial activities of water- and ethanol-extracts from Betula platyphylla var. japonica, Punica granatum and Rhus javanica. The antibacterial activity of the extracts was measured by confirming the lowest concentrations in disk diffusion and minimum inhibitory concentration (MIC) tests. Results : It was observed that RJE (Rhus javanica-EtOH extracts) show the highest content of total phenol. In addition, for RJE the total phenol content was higher from samples taken from domestic sources than from samples taken from foreign sources. DPPH radical scavenging activity was increased by treatment with PGE (Punica granatum-EtOH extracts), RJE and RJW (Rhus javanica-water extracts). It was observed that SOD-like activity was highest in the treatment with PGE. All of the extracts showed antimicrobial activity on S. epidermidis, S. aureus, P. acnes and E. coli, including those from Punica granatum, and it was noted that the activity was higher with RJE than with RJW. Conclusions : These results provide evidence that ethanol extracts of Punica granatum and Rhus javanica may have a beneficial role as antioxidants and antibiotics. Extracts from domestic samples of Betula platyphylla var japonica appeared to have a greater efficacy than extracts from foreign samples.
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